Open Access

Molecular phylogeny of Triatomini (Hemiptera: Reduviidae: Triatominae)

  • Silvia Andrade Justi1,
  • Claudia A M Russo1,
  • Jacenir Reis dos Santos Mallet2,
  • Marcos Takashi Obara3 and
  • Cleber Galvão4Email author
Parasites & Vectors20147:149

https://doi.org/10.1186/1756-3305-7-149

Received: 4 November 2013

Accepted: 15 March 2014

Published: 31 March 2014

Abstract

Background

The Triatomini and Rhodniini (Hemiptera: Reduviidae) tribes include the most diverse Chagas disease vectors; however, the phylogenetic relationships within the tribes remain obscure. This study provides the most comprehensive phylogeny of Triatomini reported to date.

Methods

The relationships between all of the Triatomini genera and representatives of the three Rhodniini species groups were examined in a novel molecular phylogenetic analysis based on the following six molecular markers: the mitochondrial 16S; Cytochrome Oxidase I and II (COI and COII) and Cytochrome B (Cyt B); and the nuclear 18S and 28S.

Results

Our results show that the Rhodnius prolixus and R. pictipes groups are more closely related to each other than to the R. pallescens group. For Triatomini, we demonstrate that the large complexes within the paraphyletic Triatoma genus are closely associated with their geographical distribution. Additionally, we observe that the divergence within the spinolai and flavida complex clades are higher than in the other Triatoma complexes.

Conclusions

We propose that the spinolai and flavida complexes should be ranked under the genera Mepraia and Nesotriatoma. Finally, we conclude that a thorough morphological investigation of the paraphyletic genera Triatoma and Panstrongylus is required to accurately assign queries to natural genera.

Keywords

Triatomini Species complex Monophyly

Background

Chagas disease, or American Trypanosomiasis, is one of the 10 most seriously neglected tropical diseases [1]. It currently affects nine million people [2], and more than 70 million people live under a serious risk of infection [3]. This vector-borne disease is transmitted by triatomine bugs (kissing bugs) infected with the parasite Trypanosoma cruzi[4]. All 148 described species of the Triatominae subfamily (Hemiptera: Reduviidae) are considered potential Chagas disease vectors [5, 6].

The Triatominae subfamily includes 15 genera, seven of which comprise the Triatomini tribe, the most diverse, and two of which are assigned to the Rhodniini tribe, the second most diverse concerning species number [6]. In the most recent taxonomic review of this group, the authors suggested synonymisation of the genera Meccus, Mepraia and Nesotriatoma with Triatoma, which is the most diverse genus of the subfamily. The generic status of these groups has been under contention because there is no consensus on whether each group constitutes a species complex or a genus [59].

The genus Triatoma is diverse in terms of the number of species (it includes 82) [6, 10, 11] and morphology. This diversity has led to the division of Triatoma into complexes based on their morphological similarities and geographic distributions [69], but no formal cladistic analysis has been performed to corroborate the assignment of these groups.

Although species complexes are not formally recognized as taxonomic ranks and, thus, do not necessarily represent natural groups, we propose that they should be monophyletic. This statement is tightly linked to the idea that once the relationships between vector species are known, information about a species may be reliably extrapolated to other closely related species [12]. Previous molecular phylogenetic studies have shown that some Triatoma complexes are not monophyletic [13, 14]. However, most of these molecular analyses were based on a single specimen per species and a single molecular marker.

The Rhodniini tribe comprises two genera: Rhodnius (18 species) and Psammolestes (three species), the former being divided into three species groups, namely, pallescens, prolixus and pictipes[15]. Although the relationship between these groups has not yet been established, with results in the literature conflicting [13, 16], it seems that Rhodnius is a paraphyletic lineage, with Psammolestes being closely related to the prolixus group [16].

In this study, we investigated which groups (genera and species complexes) within Triatomini constitute natural groups. To this end, we conducted a comprehensive molecular phylogenetic analysis of Triatomini, pioneering the inclusion of all Triatomini genera, many specimens per species and several markers per sample. We also included representatives of the three Rhodniini groups to further test ingroup monophyly. The results enabled us to accurately classify the higher groups within the Triatomini tribe, to identify monophyletic genera and complexes and to pinpoint which of these groups should be subjected to a rigorous morphological review to accurately assign natural groups.

Methods

Taxon sampling

The sampling strategy applied in this study aimed to include specimens from different populations representing the largest possible diversity of Triatomini to test the validity of current taxonomic assignments. A total of 104 specimens representing 54 Triatomini species were included, including sequences available in GenBank. To further test ingroup monophyly, we also included 10 Rhodniini species. Stenopoda sp. (Stenopodainae: Reduviidae), a member of a distinct subfamily of Reduviidae [17], was selected as the outgroup. The employed Triatominae nomenclature followed the most recently published review on the subfamily [6].

Voucher specimens for all of the adult samples sequenced in this study were deposited in the Herman Lent Triatominae Collection (CT-IOC) at the Instituto Oswaldo Cruz, FIOCRUZ. All the information about the specimens can be found in Table 1. Some of the obtained specimens consisted of first-instar nymphs, eggs or adult legs. These specimens were not deposited in the collection because the entire sample was used for DNA extraction. Nevertheless, the identification of these specimens was reliable because they were obtained from laboratory colonies with known identities of the parental generation.
Table 1

Specimens examined, including laboratory colony source, locality information (when available), voucher depository, ID (unique specimen identifier number), and GenBank accession numbers

Species

ID

Voucher number

Source

Geographic origen

Marker

COI

COII

CytB

16S

28S

18S

D. maxima

92

3465

LDP

México

KC249306

-

KC249226

KC248968

KC249134

KC249092

186

3520

LaTec

El Triunfo, México

KC249305

KC249399

KC249225

KC248967

-

-

E.mucronatus

-

-

GenBank

-

-

-

-

JQ897794

JQ897635

JQ897555

H. matsunoi

106

-

LNIRTT

 

-

KC249400

-

-

-

-

Linshcosteus sp.

-

-

GenBank

-

-

-

-

AF394595

-

-

P. geniculatus

-

-

GenBank

-

-

-

-

AF394593

-

-

P. lignarius

-

-

GenBank

-

AF449141

-

-

AY185833

-

-

P. lutzi

202

3524

LTL

Santa Quitéria, CE, Brazil

KC249307

KC249401

KC249227

KC248969

KC249135

-

P. megistus

128

3463

LACEN

Nova Prata, RS, Brazil

KC249308

KC249402

KC249228

KC248970

KC249136

-

129

3476

LACEN

Boa Vista do Cadeado, RS, Brazil

KC249309

-

KC249229

KC248971

KC249137

-

130

3477

LACEN

Tres Passos, RS, Brazil

-

-

KC249230

KC248972

KC249138

-

131

3478

LACEN

Salvador do Sul, RS, Brazil

KC249310

-

KC249231

KC248973

KC249139

-

132

3479

LACEN

Barão do Triunfo, RS, Brazil

KC249311

-

-

KC248974

KC249140

-

183

3517

LaTec

Pitangui, MG, Brazil

KC249312

KC249403

KC249232

KC248975

KC249141

-

P. tupynambai

127

3462

LACEN

Dom Feliciano, RS, Brazil

-

-

KC249233

KC248977

-

-

138

3485

LACEN

Pinheiro Machado, RS, Brazil

-

KC249404

KC249234

KC248978

KC249142

-

Paratriatoma hirsuta

-

-

GenBank

-

-

-

-

FJ230443

-

-

R. brethesi

197

3426

LNIRTT

Acará River, AM, Brazil

KC249313

KC249405

KC249235

KC248980

-

-

R. colombiensis

-

-

GenBank

-

-

-

FJ229360

AY035438

-

-

R. domesticus

-

-

GenBank

-

-

-

-

AY035440

-

-

R. ecuadoriensis

-

-

GenBank

-

-

GQ869665

-

-

-

-

R. nasutus

-

-

GenBank

-

-

-

-

-

AF435856

-

R. neivai

-

-

GenBank

-

AF449137

-

-

-

-

-

R. pallescens

-

-

GenBank

-

-

-

EF071584

-

-

-

R. pictipes

200

3429

LNIRTT

Bega, Abaetetuba, PA, Brazil

KC249315

KC249408

-

KC248982

-

KC249094

R. prolixus

-

-

GenBank

-

AF449138

-

-

-

AF435862

AY345868

R. stali

195

3424

LNIRTT

Alto Beni, Bolivia

KC249316

KC249409

KC249236

KC248983

-

-

Stenopoda sp

-

-

GenBank

-

-

-

-

FJ230414

FJ230574

FJ230493

T. brasiliensis

40

3384

LNIRTT

Curaçá, BA, Brazil

KC249319,KC249320

KC249415,KC249416

KC249240

KC248986

-

-

41

3385

LNIRTT

Sobral, CE, Brazil

-

-

KC249241

KC248987

-

-

174

3510

LaTec

Tauá, CE, Brazil

KC249318

KC249413

KC249239

KC248985

KC249145

-

T. breyeri

56

-

IIBISMED

Mataral, Cochabamba, Bolivia

KC249321

KC249417

KC249242

KC248988

-

-

T. bruneri

98

3468

LNIRTT

Cuba

-

KC249418

-

KC248989

KC249146

-

T. carcavalloi

78

3395

LNIRTT

São Gerônimo, RS, Brazil

KC249322

KC249419

KC249244

KC248991

-

KC249097

T. circummaculata

120

-

LNIRTT

Caçapava do Sul, RS, Brazil

KC249323

KC249421

-

KC248992

KC249147

KC249098

121

-

LACEN

Piratini, RS, Brazil

KC249324

KC249422

-

KC248993

-

-

122

3473

LACEN

Piratini, RS, Brazil

KC249325

-

KC249245

KC248994

KC249148

KC249099

126

3461

LACEN

Dom Feliciano, RS, Brazil

-

-

-

KC248996

-

-

T. costalimai

35

3381

LNIRTT

Posse, GO, Brazil

KC249327,KC249328

KC249425

KC249246

KC248997

-

KC249101

42

-

IIBISMED

Chiquitania, Cochabamba, Bolivia

KC249329

KC249426

KC249247

KC248998

KC249149

-

T. delpontei

53

-

IIBISMED

Chaco Tita, Cochabamba, Bolivia

KC249330

KC249427

KC249248

KC248999

-

-

T. dimidiata

20

3444

LaTec

-

KC249335

KC249431

-

KC249004

KC249152

-

94

3466

LNIRTT

Central América

KC249336,KC249337

KC249432

-

KC249005

KC249155

-

100

3470

LNIRTT

México

KC249333

-

-

KC249002

-

-

T. eratyrusiformis

-

-

GenBank

-

GQ336898

-

JN102360

AY035466

-

-

T. flavida

-

-

GenBank

-

-

-

-

AY035451

-

AJ421959

T. garciabesi

89

3405

LNIRTT

Rivadaria, Argentina

KC249338

-

KC249249

KC249006

KC249158

KC249102

T. guasayana

55

-

IIBISMED

Chaco Tita, Cochabamba, Bolivia

KC249342

-

KC249251

KC249010

-

-

82

3398

LNIRTT

Santa Cruz, Bolívia

KC249343

KC249438

KC249252

KC249011

KC249162

KC249103

T. guazu

29

3455

LNIRTT

Barra do Garça, MT, Brazil

-

KC249440

-

KC249013

KC249164

KC249105

T. infestans

58

-

IIBISMED

Cotapachi, Cochabamba, Bolivia

KC249349

KC249442

KC249256

KC249015

KC249168

KC249109

60

-

IIBISMED

Mataral, Cochabamba, Bolivia

KC249351

KC249443

KC249257

KC249016

KC249169

KC249107

62

-

IIBISMED

Ilicuni, Cochabamba, Bolivia

KC249353

KC249445

KC249259

KC249018

-

-

63

-

IIBISMED

Ilicuni, Cochabamba, Bolivia

KC249354

KC249446

KC249260

KC249019

-

-

66

3386

LNIRTT

Guarani das Missões, RS, Brazil

-

-

-

KC249021

-

-

68

3388

LNIRTT

Argentina

-

-

-

KC249023

-

-

69

3389

LNIRTT

Montevideo, Uruguai

-

KC249447

KC249262

KC249024

KC249172

-

44

-

IIBISMED

Chaco Tita Cochabamba

KC249346

-

KC249255

KC249025

KC249166

KC249108

T. juazeirensis

209

3430

LTL

Uiabí, BA, Brazil

-

-

KC249263

KC249026

KC249173

-

T. jurbergi

30

3456

LNIRTT

Alto Garça MT, Brazil

-

KC249448

KC249264

KC249027

KC249174

KC249110

T. klugi

75

3393

LNIRTT

Nova Petrópolis, RS, Brazil

KC249356

KC249449

KC249265

KC249028

-

-

T. lecticularia

151

3411

LaTec

-

-

KC249450

-

KC249029

KC249175

KC249111

T. longipennis

26

3450

LaTec

-

-

KC249453

KC249267

KC249032

-

-

97

3467

LNIRTT

México

KC249358

-

-

KC249033

-

-

165

3501

LaTec

México

KC249357

KC249452

-

KC249031

KC249177

-

T. maculata

203

3525

LTL

Água Fria, RR, Brazil

-

KC249454

-

KC249034

 

-

T. matogrossensis

31

3374

LNIRTT

Bahia, Brazil

KC249361

KC249458

-

KC249038

-

-

32

3375

LNIRTT

Aquidauana , MS, Brazil

-

KC249459

KC249271

KC249039

KC249181

-

33

3377

LNIRTT

Alegria, MT, Brazil

-

KC249460

KC249272

KC249040

KC249182

KC249114

192

3423

LTL

São Gabriel D’oeste, MS, Brazil

KC249360

KC249457

KC249270

KC249037

KC249180

KC249113

T. mazzottii

-

-

GenBank

-

DQ198805

-

DQ198816

AY035446

-

AJ243333

T. melanica

-

3447

LaTec

-

-

KC249461

-

KC249041

KC249183

-

T. melanosoma

70

3390

LNIRTT

Missiones Argentina

KC249362

-

KC249273

KC249042

-

-

T. nitida

-

-

GenBank

-

-

-

AF045723

AF045702

-

-

T. pallidipennis

18

3442

LaTec

-

-

-

-

KC249045

-

-

T. phyllosoma

-

-

GenBank

-

DQ198806

-

DQ198818

-

-

AJ243329

T. picturata

-

-

GenBank

-

-

-

DQ198817

AY185840

-

AJ243332

T. platensis

96

-

LNIRTT

Montevideo Uruguai

-

-

KC249274

KC249047

KC249186

-

T. protracta

93

3407

LNIRTT

Monte Diablo, California, EUA

-

KC249463

-

KC249048

KC249187

-

T. pseudomaculata

34

3379

LNIRTT

Curaçá, BA, Brazil

-

-

-

KC249057

KC249196

-

211

3432

LTL

Várzea Alegre, CE, Brazil

KC249364

KC249464

KC249275

KC249050

KC249189

-

212

3433

LTL

Várzea Alegre, CE, Brazil

-

KC249465

KC249276

KC249051

KC249190

-

214

3435

LTL

Várzea Alegre, CE, Brazil

KC249365

KC249467

KC249277

KC249053

KC249192

-

T. recurva

-

-

GenBank

-

DQ198803

-

DQ198813

FJ230417

-

FJ230496

T. rubrofasciata

-

-

GenBank

-

-

-

-

AY127046

-

AJ421960

T. rubrovaria

76

3459

LNIRTT

Caçapava do Sul, RS, Brazil

KC249375

KC249477

KC249286

KC249066

-

-

77

3394

LNIRTT

Quevedos, RS, Brazil

KC249376

-

KC249287

KC249067

KC249204

KC249122

156

3416

LaTec

Canguçu, RS, Brazil

KC249374

KC249476

KC249285

KC249065

KC249203

KC249121

123

3474

LACEN

Piratini, RS, Brazil

KC249369

KC249470

-

KC249058

KC249197

KC249116

134

3481

LACEN

Canguçu, RS, Brazil

KC249370

KC249471

KC249281

KC249059

KC249198

KC249117

136

3483

LACEN

Pinheiro Machado, RS, Brazil

KC249372

KC249473

KC249283

KC249061

KC249200

KC249119

140

3487

LACEN

Canguçu, RS, Brazil

KC249373

KC249475

-

KC249064

KC249202

KC249120

T. sanguisuga

-

-

GenBank

-

-

JF500886

HQ141317|

AF045696

-

-

T. sherlocki

80

3396

LNIRTT

-

KC249377

KC249478

KC249288

KC249068

KC249205

-

T. sordida

38

3382

LNIRTT

Rondonópolis, MT, Brazil

-

KC249479

-

KC249071

-

-

46

-

IIBISMED

Romerillo, Cochabamba, Bolivia

KC249379,KC249380

KC249480

-

KC249072

KC249207

-

47

-

IIBISMED

Romerillo, Cochabamba, Bolivia

KC249381,KC249382

-

KC249290

KC249073

KC249208

KC249124

83

3399

LNIRTT

La Paz, Bolívia

KC249383

KC249481

KC249291

KC249074

KC249209

-

85

3401

LNIRTT

Pantanal, MS, Brazil

KC249384

KC249482

KC249292

KC249075

KC249210

KC249125

86

3402

LNIRTT

Santa Cruz, Bolívia

KC249385

-

KC249293

KC249076

KC249211

-

88

3404

LNIRTT

San Miguel Corrientes, Argentina

KC249387

KC249484

KC249295

KC249078

KC249213

-

90

3406

LNIRTT

Poconé, MT, Brazil

KC249388

-

-

KC249079

-

-

Triatoma sp.

50

-

IIBISMED

Mataral, Cochabamba, Bolivia

KC249339

KC249435

-

KC249007

KC249159

-

T. spinolai

-

-

GenBank

-

GQ336893

-

JN102358

AF324518

-

AJ421961

T. tibiamaculata

79

3460

LNIRTT

-

KC249390

KC249486

KC249297

KC249081

KC249215

-

177

3513

LaTec

Mogiguaçu, RS, Brazil

KC249389

KC249485

KC249296

KC249080

KC249214

KC249127

T. vandae

28

3452

LNIRTT

Pantanal, MT, Brazil

KC249391

KC249487

KC249298

KC249082

KC249216

KC249128

73

3392

LNIRTT

Rio Verde do Mato Grosso, MT, Brazil

KC249392

KC249488

KC249299

KC249083

KC249217

KC249129

74

3458

LNIRTT

Rondonópolis, MT, Brazil

KC249393,KC249394

KC249489

KC249300

KC249084

KC249218

-

T. vitticeps

81

3397

LNIRTT

-

KC249396

KC249491

KC249303

KC249087

KC249220

KC249132

91

-

LTL

Rio de Janeiro, Brazil

KC249397

KC249492

KC249304

KC249088

KC249221

-

168

3504

LaTec

Itanhomi, MG, Brazil

KC249395

KC249490

KC249301

KC249085

-

KC249130

T. williami

36

-

LNIRTT

-

-

KC249493

-

KC249089

-

-

T. wygodzynski

17

3441

LaTec

-

KC249398

KC249494

-

KC249090

KC249222

KC249133

205

3527

LTL

Sta Rita de Caldas, MG, Brazil

-

-

-

KC249091

-

-

LTL - Laboratório de Transmissores de Leishmanioses, IOC, FIOCRUZ; LaTec - Laboratório de Triatomíneos e epidemiologia da Doença de Chagas, CPqRR, FIOCRUZ; LACEN - Laboratório Central, Rio Grande do Sul, Ministério da Saúde; IIBISMED - Instituto de Investigaciones Biomédicas, Facultad de Medicina, Universidad Mayor de San Simón, Cochabamba, Bolivia.

DNA extraction, amplification and sequencing

The DNA extraction was performed using the protocol described by Aljanabi and Martinez [18] or using the Qiagen Blood and Tissue kit, according to the manufacturer’s recommendations. The following PCR cycling conditions were employed: 95°C for 5 min; 35 cycles of 95°C for 1 min, 49–45°C for 1 min, and 72°C for 1 min; and 72°C for 10 min. The sequences of the primers used for amplification are shown in Table 2. The reaction mixtures contained 10 mM Tris–HCl/50 mM KCl buffer, 0.25 mM dNTPs, 10 μM forward primer, 10 μM reverse primer, 3 mM MgCl2, 2.5 U of Taq polymerase and 10–30 ng of DNA. The primers used to amplify the mitochondrial COI, COII, CytB and 16S and the nuclear ribosomal 18S and 28S markers are listed in Table 2.
Table 2

Primers used in this study

Marker

Forward primer

Reverse primer

COI

5′-GGTCAACAAATCATAAAGATATTGG-3′ [19]

5′-AAACTTCAGGGTGACCAAAAAATCA-3′ [19]

5′-CCTGCAGGAGGAGGAGAYCC-3′ [20]

5′ - TAAGCGTCTGGGTAGTCTGARTAKCG-3′; [21]

5′-ATTGRATTTTDAGTCATAGGGAG-3′ (this study)

5′-TATTYGTWTGATCDGTWGG-3′ (this study)

CytB

5′-GGACG(AT)GG(AT)ATTTATTATGGATC-3′ [22]

5′-ATTACTCCTCCTAGYTTATTAGGAATT-3′ [23]

COII

5′-ATGATTTTAAGCTTCATTTATAAAGAT-3′ [23]

5′-GTCTGAATATCATATCTTCAATATCA-3′ [23]

16S

5′-CGCCTGTTTATCAAAAACAT-3′ [24]

5′-CTCCGGTTTGAACTCAGATCA-3′ [24]

28S

5′- AGTCGKGTTGCTTGAKAGTGCAG-3′ [25]

5′- TTCAATTTCATTKCGCCTT-3′ [25]

 

5′-CTTTTAAATGATTTGAGATGGCCTC-3′ (this study)

-

18S

5′-AAATTACCCACTCCCGGCA-3′ [24]

5′-TGGTGUGGTTTCCCGTGT T-3′ [24]

The PCR-amplified products were purified using the ExoSAP-IT (USB® products), according to the manufacturer’s recommendations, and both strands were subsequently sequenced. The sequencing reactions were performed using the ABI Prism® BigDye® Terminator v3.1 Cycle Sequencing kit (Applied Biosystems), with the same primers employed for PCR, in ABI 3130 and ABI 3730 sequencers (PDTIS Platform, FIOCRUZ and the Genetics Department of UFRJ, respectively). The obtained sequences were assembled using MEGA 4.0 [26] and SeqMan Lasergene v. 7.0 (DNAStar, Inc.) software.

Sequence alignments and molecular datasets

Different approaches were used to align the coding sequences and the ribosomal DNA markers. The coding sequences were translated and then aligned using ClustalW [27] implemented in MEGA 4.0 [26] software. The ribosomal DNA sequences were aligned using MAFFT [28] with the Q-INS-I option, which takes the secondary RNA structure into consideration.

We first constructed an alignment including all the sequences obtained (Additional file 1: Table S1), but there was too much missing data in this matrix, which included 169 individuals. To minimise the effect of missing data on the analysis, a new alignment was constructed based on the above method with the aim of maximising diversity, considering that each taxon in the dataset had to be comparable to all others, that is, all specimens must include comparable sequences.

The final individual alignments were concatenated by name using SeaView [29], generating a matrix including 115 individuals and 6,029 nucleotides (Table 1). This dataset is available on the Dryad database (http://datadryad.org/) and upon request.

Phylogenetic analyses

jModeltest [30] was used to assess the best fit model for each of the markers. The markers CytB, COII, 18S and 28S fit models less parametric than GTR + Γ (data not shown). Despite this fact, GTR + Γ was used for all the markers as this is the next best model available in the programs used. The use of a more parametric model is supported by the fact that the application of a model less parametric than the “real” model leads to a strong accentuation of errors in the recovered tree [31].

The Maximum Likelihood (ML) tree was obtained through a search of 200 independent runs with independent parsimony starting trees using RAxML 7.0.4 [32]. The alignment was partitioned by marker, and for each partition, the gamma parameter was estimated individually, coupled to the GTR model. To assess the reliability of the recovered clades, 1,000 bootstrap [33] replicates were performed using the rapid bootstrap algorithm implemented in RaxML.

Additionally, a Bayesian approach was applied to reconstruct the phylogeny of the concatenated dataset using MrBayes 3 [34]. The data were also partitioned based on markers, and GTR + Γ (four categories) was used separately for each partition, with the gamma parameter being estimated individually. The trees were sampled every 1,000 generations for 100 million generations in two independent runs with four chains each. Burn-in was set to 50% of the sampled trees.

Results

The recovered phylogenies (ML and BI) yielded very similar trees, with the generated clades supporting their agreement with one another.

The Rhodniini tribe

The Rhodniini tribe (Figure 1, Additional file 2: Figure S1 and Additional file 3: Figure S2) was recovered with high support (BS = 100, PP = 1), as were most relationships within the tribe. The prolixus group was recovered as a sister taxon to the pictipes group (BS = 97, PP = 1), and these groups form a sister clade to the pallescens group. The only species that could not be confidently placed within its clade was R. neivai, which was recovered within the prolixus group as a sister species to R. nasutus, but support was lower (BS = 80, PP = 0.7; Figure 1, Additional file 2: Figure S1 and Additional file 3: Figure S2).
Figure 1

Best ML tree (on the left) and Bayesian consensus tree (on the right) reconstructed. Bars on the right highlight the non-monophyletic groups. Numbers above branches represent clade support higher than 50 and 0.5, respectively. Triatominae photos by Carolina Dale. Stenopoda spinulosa photo by Brad Barnd. Photos are not to scale.

The Triatomini tribe

The Triatomini tribe was recovered with the highest support (BS = 100, PP = 1). The tribe was shown to be divided into three main lineages: Clade (1), Panstrongylus + the flavida complex (Nesotriatoma) + T. tibiamaculata (BS = 69, PP = 1); Clade (2), the monotypic genera (Hermanlentia, Paratriatoma, Dipetalogaster) + Linshcosteus + Northern Hemisphere Triatoma (BS = 88, PP = 1); and Clade (3), Southern Hemisphere Triatoma (including the spinolai complex or Mepraia) and Eratyrus (BS = 65, PP = 0.68).

Clade (1): Panstrongylus + the flavida complex (Nesotriatoma) + T. tibiamaculata

The flavida complex (Nesotriatoma) was recovered with the highest support in all three phylogenies, showing a close relationship with the clade formed by P. geniculatus + P. lutzi + P. tupynambai. P. megistus was placed as a sister taxon to T. tibiamaculata (BS = 95, PP = 1); while P. lignarius could not be confidently placed in the clade (BS < 50, PP = 0.55).

Clade (2): the monotypic genera (Hermanlentia, Paratriatoma, Dipetalogaster) + Linshcosteus + Northern Hemisphere Triatoma

In this clade, the phylogenies showed close relationships among Paratriatoma (Pa.), Dipetalogaster and T. nitida, T. protracta (protracta complex) and T. lecticularia (lecticularia complex). Pa. hirsuta was always recovered as a sister species to T. lecticularia (BS = 75, PP = 0.98), and this pair was sister to D. maxima (BS = 80, PP = 0.99). The indicated species from the protracta complex were always recovered as a single clade that was closely related to D. maxima, Pa. hirsuta and T. lecticularia.

The tropicopolitan T. rubrofasciata species was recovered as a sister species to Linshcosteus in both phylogenies with high support (BS = 98, PP = 1). This pair of species is closely related to the clade formed by the dimidiata subcomplex + T. sanguisuga (lecticularia subcomplex) + Hermanlentia matsunoi + the phyllosoma subcomplex + T. recurva (BS = 100, PP = 1).

H. matsunoi appeared as a sister taxon to T. dimidiata from Mexico with high support (BS = 99, PP = 1). The phyllosoma subcomplex was not recovered as monophyletic as T. recurva was recovered close to T. longipennis, although the bootstrap for this clade was not high (BS = 72, PP = 0.98).

Clade (3) Southern Hemisphere Triatoma and Eratyrus

This clade was formed by the spinolai complex and the species assigned to the infestans complex, which were not recovered as monophyletic. The spinolai complex was recovered as monophyletic (BS = 100, PP = 1) in both phylogenies, and as sister taxa to the infestans complex.

T. vitticeps was recovered as a sister taxon to E. mucronatus and to the remaining Southern Hemisphere Triatoma subcomplexes of the infestans (BS = 94, PP = 1). The infestans and rubrovaria subcomplexes were recovered as monophyletic (BS = 99, PP = 1 and BS = 83, PP = 0.99, respectively). In addition, the rubrovaria subcomplex was closely related to a short-winged Triatoma sp. (BS = 99, PP = 1) that resembles T. guasayana, which was discovered in the bromeliads of the Bolivian Chaco by F. Noireau.

T. maculata was not closely related to the other species of the maculata subcomplex. This taxon clustered with the brasiliensis subcomplex (BS = 51, PP = 0.82), except for T. tibiamaculata and T. vitticeps, which clustered elsewhere. The remaining species of the maculata subcomplex clustered in a large clade with the sordida and matogrossensis subcomplexes (BS = 71, PP = 0.98).

Discussion

Phylogenetic analyses

The reconstructed phylogenies presented in this report showed similar topologies and consistent branch support values. The posterior probability values were almost always higher than the bootstrap values, as expected [31] (Additional file 2: Figure S1 and Additional file 3: Figure S2).

Nonetheless, deep relationships, such as those between complexes, could be resolved. In addition, relationships within the infestans subcomplexes remain unclear (Additional file 2: Figure S1 and Additional file 3: Figure S2). The short terminal branches of these subcomplexes indicate that their diversification must have occurred recently. Under this scenario, incomplete lineage sorting would account for the lack of phylogenetic resolution within the group [35].

A different approach will be adopted in future studies to assess the relationships between closely related species that could not be resolved here. New unlinked nuclear markers, especially those linked to development and reproduction [36], will be sequenced to generate a species tree reconstruction [37], which is a more suitable method of phylogenetic reconstruction for closely related species.

The Rhodniini tribe

The Rhodniini tribe comprises only 2 genera: Rhodnius and Psammolestes. Rhodnius has long been known to be easily distinguishable from other Triatominae, but the morphological discrimination of the species within Rhodnius is rather difficult [38]. Moreover, there is no uncertainty in the literature regarding the species groups assigned within Rhodnius; the uncertainty is related to the relationships between these groups.

Previously described molecular phylogenies of these genera have yielded distinct results. For instance, Lyman et al.[16] showed the pallescens group to be more closely related to the pictipes group, but Hypsa et al.[13] found the pictipes group to be closer to the prolixus group, which is consistent with our results. This difference could be due to differences in taxon sampling rather than differences in the gene trees, as both of these authors used mitochondrial markers. In this work, the taxon sampling process included a larger number of species than were included by Lyman et al.[16] (see also [13]). Wiens and Tiu [39] demonstrated that the addition of taxa should improve the accuracy of a phylogenetic reconstruction. The amount of data (less than 10% of the size of our alignment) from Hypsa et al.[13] was overturned by their taxon sampling, which included twice the number of species as the first work.

The Triatomini tribe

The Triatomini tribe is the most diverse tribe within the subfamily, and many taxonomic proposals have been put forth for the groups belonging to this tribe. The most prominent of these proposals is that Meccus, Mepraia and Nesotriatoma be considered as genera or species complexes belonging to Triatoma[5, 6, 8]. Dujardin et al.[40] noted these confusing systematics with another example: the number of monotypic genera within the tribe and the number of subspecies (at times also considered separate species) assigned to Triatomini. Figure 1, based on our results, highlights the most accepted Triatomini groups that are not monophyletic. We show that Triatoma and Panstrongylus are not natural groups. However, diversities formerly placed under the generic names Mepraia and Nesotriatoma, but not Meccus, consist of monophyletic lineages.

Therefore, based on our results, we indicate that Mepraia and Nesotriatoma should be ranked as genera, as previously proposed [5]. The branch lengths of the reconstructed phylogenies (Figure 1, Additional file 2: Figure S1 and Additional file 3: Figure S2) showed much greater distances between the species assigned to each of these genera than within the other Triatoma complexes. In addition, if the species belonging to Nesotriatoma are considered a species complex of another genus, it is reasonable to include these species in the genus Panstrongylus.

Previous studies have indicated a putative paraphyletic status for Panstrongylus, despite a lack of resolution in some groups [13, 14, 41, 42]. In our topology, Panstrongylus is clearly divided into two groups: one including P. tupynambai, P. lutzi and P. geniculatus as sister taxa to Nesotriatoma and another group showing a close and highly supported relationship between T. tibiamaculata and P. megistus.

The most prominent morphological characteristic that separates Panstrongylus from other Triatomini is the short head of these species, with antennae close to the eyes [8]. The non-monophyletic status of Panstrongylus (Figure 1; see also [14]) indicates that this putative diagnostic characteristic of the genus might be a morphological convergence. Indeed, some Panstrongylus populations show variation in eye size according to their habitat, and this variation influences the distances between the antennae and the eyes [43]. Panstrongylus species tend to present Triatoma-like head shapes [43] during development when the nymphs exhibit smaller eyes. Furthermore, North American Triatoma may display smaller heads and antennae that are closer to the eyes than their South American counterparts[6].

Triatoma is composed of two distinct paraphyletic groups: one occurring in the Northern hemisphere and the other in the Southern Hemisphere; one exception found in the present work was T. tibiamaculata, which clusters with Panstrongylus elsewhere. The previous assignments of Triatoma species into complexes took into consideration the geographical distributions of the groups and their morphological features (e.g. [6]). Our results clearly indicate that monophyletic clades of Triatoma species, which do not necessarily correspond to these complexes, are correlated with restricted geographical distributions corresponding to different biogeographical provinces [44]. This is particularly evident in South America.

Northern Hemisphere Triatoma and the less diverse genera

T. lecticularia is sister to Pa. hirsuta. This pair of species is closely related to D. maxima, which is a genus whose head shape resembles a large Triatoma. Furthermore, Pa. hirsuta exhibits a head shape similar to T. lecticularia, which was observed by Lent and Wygodzinsky [8].

H. matsunoi, which was included in a phylogenetic study for the first time in the present work, was recovered as the sister taxon to the Mexican lineage of T. dimidiata. H. matsunoi was first described as belonging to Triatoma[45] based on the main features used to characterise the Triatomini genera. Subsequently, Jurberg and Galvão [46] found major differences in the male genitalia of this species relative to other Triatomini and reassigned it to a new monotypic genus.

T. rubrofasciata appears to be the species that is closest to Linshcosteus, which is the only Triatomini genus exclusively from the Old World, more precisely, from India. Although we did not include Old World Triatoma in our analyses, previous morphometric analyses have shown Linshcosteus to be distinct from Old world Triatoma and from the closely related species T. rubrofasciata from the New World [47].

The dimidiata subcomplex was not recovered as a natural group because the two sampled T. dimidiata s.l. lineages [48] did not cluster, and the clade also included T. lecticularia and H. matsunoi. Consistent with our results, Espinoza et al.[49] recently published a reconstructed phylogeny showing the relationships among the North American Triatoma species. They included T. gerstaeckeri and T. brailovskyi (not included here) in their analysis and demonstrated the close relationships between these species and those from the dimidiata and phyllosoma subcomplexes, confirming the need to review these groups.

Southern Hemisphere Triatoma

Most subcomplexes assigned to the infestans complex were not recovered as monophyletic. The only natural groups recovered were the infestans and rubrovaria subcomplexes.

As noted above, most of the monophyletic clades recovered for these Triatoma can be associated with a South American biogeographical province. This shows that geographical distribution currently has greater importance than morphology in the process of assigning natural groups to the genus. Henceforth, the geographical provinces (related to biomes) will be referred to as described in Morrone [44].

T. vitticeps, the first Triatoma lineage to diverge in this clade, is found in the Atlantic Forest and shares morphological similarities with the unsampled species T. melanocephala[6], which is a rare species found exclusively in northeastern Brazil [50]. Although both species were assigned to the former brasiliensis complex ([6]), both our results and the number of sex chromosomes in these species, which differs from the other Southern Hemisphere Triatoma, would exclude them from this group [50].

The next lineage to diverge in this clade was Eratyrus mucronatus. The genus Eratyrus differs from Triatoma in displaying a long spine-shaped posterior process of the scutellum and a long first rostral segment, which is nearly as long as the second segment [8]. Although we did not include E. cuspidatus in our analysis, the morphology of this genus is rather distinct, and apart from its phylogenetic position within Triatoma, this species is not a subject of “systematic dispute” in the literature.

Triatoma maculata appears as the sister taxon to part of the brasiliensis subcomplex (except T. tibiamaculata and T. vitticeps). Previous studies have demonstrated the close relationships among some species in the brasiliensis subcomplex [51]. However, these studies did not include T. maculata in their analyses. In contrast, an earlier study revealed a possible close relationship between T. brasiliensis and T. maculata[13]. T. maculata is exclusively found in the Amazonian forest, while the brasiliensis subcomplex is exclusive to the Caatinga province in northeastern Brazil.

The species assigned to the infestans, sordida, and rubrovaria subcomplexes currently exhibit overlapping distributions as they all occur in the Chacoan dominion. The infestans subcomplex was found to be monophyletic, with its distribution occurring mainly in Chaco province. It is important to highlight that only sylvatic populations were considered for this designation because T. infestans shows a distribution related to human migration in most Southern American countries [52].

The Triatoma sp. informally described by François Noireau as a short-winged form of T. guasayana appears as the sister taxon to the rubrovaria subcomplex. This previously undescribed species was collected in Chaco province from bromeliads, which form a different microhabitat than the rock piles in which rubrovaria species are usually found [53]. Conversely, the rubrovaria subcomplex is restricted to Pampa province and the Paraná dominion. As Pampa and Chaco provinces belong to the Chacoan dominion, Triatoma sp. and the rubrovaria complex inhabit historically related areas [44], we predict that microhabitat adaptations account for the morphological divergence observed between these groups.

The most morphologically diverse clade includes species from the sordida, maculata (except for T. maculata) and matogrossensis subcomplexes. This is also the most widespread group in South America and occupies most of Cerrado and Chaco provinces.

Conclusions

Our results show that a thorough evolutionary mapping of the morphological characteristics of Triatomini is long overdue. For example, head shape, which was previously used to distinguish Panstrongylus from Triatoma, does not appear to be a reliable characteristic; the highly supported P. megistus + T. tibiamaculata sister taxa corroborate this conclusion.

In addition, the only published cladistic analysis of a Triatominae group, for Panstrongylus[8], does not agree with our results, though this might be due to the fact that Nesotriatoma and T. tibiamaculata were not included in their analysis. We have shown that the genus Triatoma and a majority of the Triatoma species complexes are not monophyletic. Knowledge of morphologies and the evolutionary histories of morphological traits are imperative in assigning natural groups. In the case of Triatomini, such knowledge is particularly relevant due to the epidemiological importance of these organisms [12].

Declarations

Acknowledgements

We thank L. Diotaiuti (LaTec, CPqRR, FIOCRUZ) and A.C.V. Junqueira (LDP, IOC, FIOCRUZ) for samples, C. Dale (LNIRTT) and A.E.R. Soares (LBETA) for reviewing the manuscript and PDTIS-FIOCRUZ for the use of their DNA sequencing facilities. SA Justi is supported by a scholarship from the Brazilian National Research Council (CNPq), and this study is part of the requirement for her doctorate at the Federal University of Rio de Janeiro under the supervision of CAM Russo. We dedicate this study to the memory of Dr. François Noireau.

Authors’ Affiliations

(1)
Departamento de Genética, Laboratório de Biologia Evolutiva Teórica e Aplicada, Universidade Federal do Rio de Janeiro, CCS, Instituto de Biologia
(2)
Laboratório de Transmissores de Leishmanioses, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz
(3)
Faculdade de Ceilândia, Universidade de Brasília
(4)
Laboratório Nacional e Internacional de Referência em Taxonomia de Triatomíneos, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz

References

  1. World Health Organization: WHO Roadmap Inspires Unprecedented Support to Defeat Neglected Tropical Diseases. 2012, Geneva, Switzerland: World Health OrganizationGoogle Scholar
  2. Schofield CJ, Jannin J, Salvatella R: The future of Chagas disease control. Trends Parasitol. 2006, 22: 583-588. 10.1016/j.pt.2006.09.011.View ArticlePubMedGoogle Scholar
  3. Coura JR: Chagas disease: what is known and what is needed – a background article. Mem I Oswaldo Cruz. 2007, 102: 113-122.Google Scholar
  4. Chagas CRJ: Nova tripanozomiaze humana: estudos sobre a morfolojia e o ciclo evolutivo do Schizotrypanum cruzi n. gen, n. sp, ajente etiolojico de nova entidade morbida do homem. Mem I Oswaldo Cruz. 1909, 1: 159-218. 10.1590/S0074-02761909000200008.View ArticleGoogle Scholar
  5. Galvão C, Carcavallo R, Rocha DS, Jurberg J: A checklist of the current valid species of the subfamily Triatominae Jeannel, 1919 (Hemiptera, Reduviidae) and their geographical distribution, with nomenclatural and taxonomic notes. Zootaxa. 2003, 202: 1-36.Google Scholar
  6. Schofield CJ, Galvão C: Classification, evolution, and species groups within the Triatominae. Acta Trop. 2009, 110: 88-100. 10.1016/j.actatropica.2009.01.010.View ArticlePubMedGoogle Scholar
  7. Usinger RL, Wygodzinsky P, Ryckman RE: The biosystematics of Triatominae. Ann Rev Entomol. 1966, 11: 309-330. 10.1146/annurev.en.11.010166.001521.View ArticleGoogle Scholar
  8. Lent H, Wygodzinsky P: Revision of the Triatominae (Hemiptera: Reduviidae) and their significance as vectors of Chagas disease. B Am Mus Nat Hist. 1979, 163: 123-520.Google Scholar
  9. Carcavallo RU, Jurberg J, Lent H, Noireau F, Galvão C: Phylogeny of the Triatominae (Hemiptera: Reduviidae). Proposals for taxonomic arrangements. Entomol Vect. 2000, 7 (Supp l1): 1-99.Google Scholar
  10. Gonçalves TCM, Teves-Neves SC, Santos-Mallet JR, Carbajal-de-la-Fuente AL, Lopes CM: Triatoma jatai sp. nov. in the state of Tocantins, Brazil (Hemiptera: Reduviidae: Triatominae). Mem I Oswaldo Cruz. 2013, 108: 429-437.View ArticleGoogle Scholar
  11. Jurberg J, Cunha V, Cailleaux S, Raigorodschi R, Lima MS, Rocha DS, Moreira FFF: Triatoma pintodiasi sp. nov. do subcomplexo T. rubrovaria (Hemiptera, Reduviidae, Triatominae). Rev Pan-Amaz Saúde. 2013, 4: 43-56.View ArticleGoogle Scholar
  12. Schaefer CW: Triatominae (Hemiptera: Reduviidae): systematic questions and some others. Neotrop Entomol. 2003, 32: 1-10. 10.1590/S1519-566X2003000100001.View ArticleGoogle Scholar
  13. Hypsa V, Tietz DF, Zrzavý J, Rego RO, Galvão C, Jurberg J: Phylogeny and biogeography of Triatominae (Hemiptera: Reduviidae): molecular evidence of a New World origin of the Asiatic clade. Mol Phylogenet Evol. 2002, 23: 447-457. 10.1016/S1055-7903(02)00023-4.View ArticlePubMedGoogle Scholar
  14. Marcilla A, Bargues MD, Abad-Franch F, Panzera F, Carcavallo RU, Noireau F, Galvão C, Jurberg J, Miles MA, Dujardin JP, Mas-Coma S: Nuclear rDNA ITS-2 sequences reveal polyphyly of Panstrongylus species (Hemiptera: Reduviidae: Triatominae), vectors of Trypanosoma cruzi. Infect Genet Evol. 2002, 1: 225-235. 10.1016/S1567-1348(02)00029-1.View ArticlePubMedGoogle Scholar
  15. Abad-Franch F, Monteiro FA, Jaramillo ON, Gurgel-Gonçalves R, Dias FB, Diotaiuti L: Ecology, evolution, and the long-term surveillance of vector-borne Chagas disease: a multi-scale appraisal of the tribe Rhodniini (Triatominae). Acta Trop. 2009, 110: 159-177. 10.1016/j.actatropica.2008.06.005.View ArticlePubMedGoogle Scholar
  16. Lyman DF, Monteiro FA, Escalante AA, Cordon-Rosales C, Wesson DM, Dujardin JP, Beard CB: Mitochondrial DNA sequence variation among triatomine vectors of Chagas’ disease. Am J Trop Med Hyg. 1999, 60: 377-386.PubMedGoogle Scholar
  17. Weirauch C: Cladistic analysis of Reduviidae (Heteroptera:Cimicomorpha) based on morphological characters. Syst Entomol. 2008, 33: 229-274. 10.1111/j.1365-3113.2007.00417.x.View ArticleGoogle Scholar
  18. Aljanabi SM, Martinez I: Universal and rapid salt-extraction of high quality genomic DNA for PCR based techniques. Nucleic Acids Res. 1997, 25: 4692-4693. 10.1093/nar/25.22.4692.PubMed CentralView ArticlePubMedGoogle Scholar
  19. Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R: DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol Mar Biol Botech. 1994, 3: 294-299.Google Scholar
  20. Palumbi SR, Benzie J: Large mitochondrial DNA differences between morphologically similar Penaeid shrimp. Mol Mar Biol Biotech. 1991, 1: 27-34.Google Scholar
  21. Baldwin JD, Bass AL, Bowen BW, Clark WH: Molecular phylogeny and biogeog- raphy of the marine shrimp Penaeus. Mol Phylogenet Evol. 1998, 10: 399-407. 10.1006/mpev.1998.0537.View ArticlePubMedGoogle Scholar
  22. Monteiro FA, Barrett TV, Fitzpatrick S, Cordon-Rosales C, Feliciangeli D, Beard CB: Molecular phylogeography of the Amazonian Chagas disease vectors Rhodnius prolixus and R. robustus. Mol Ecol. 2003, 12: 997-1006. 10.1046/j.1365-294X.2003.01802.x.View ArticlePubMedGoogle Scholar
  23. Patterson PS, Gaunt MW: Phylogenetic multilocus codon models and molecular clocks reveal the monophyly of haematophagous reduviid bugs and their evolution at the formation of South America. Mol Phyl Evol. 2010, 56: 608-621. 10.1016/j.ympev.2010.04.038.View ArticleGoogle Scholar
  24. Weirauch C, Munro JB: Molecular phylogeny of the assassin bugs (Hemiptera: Reduviidae), based on mitochondrial and nuclear ribosomal genes. Mol Phylogenet Evol. 2009, 53: 287-299. 10.1016/j.ympev.2009.05.039.View ArticlePubMedGoogle Scholar
  25. Dietrich CH, Rakitov RA, Holmes JL, Black WC: Phylogeny of the major lineages of Membracoidea (Insecta: Hemiptera: Cicadomorpha) based on 28S rDNA sequences. Mol Phylogenet Evol. 2001, 18: 293-305. 10.1006/mpev.2000.0873.View ArticlePubMedGoogle Scholar
  26. Tamura K, Dudley J, Nei M, Kumar S: MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol. 2007, 24: 1596-1599. 10.1093/molbev/msm092.View ArticlePubMedGoogle Scholar
  27. Thompson JD, Higgins DG, Gibson TJ: CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994, 22: 4673-4680. 10.1093/nar/22.22.4673.PubMed CentralView ArticlePubMedGoogle Scholar
  28. Katoh K, Toh H: Improved accuracy of multiple ncRNA alignment by incorporating structural information into a MAFFT-based framework. BMC Bioinformatics. 2008, 9: 212-10.1186/1471-2105-9-212.PubMed CentralView ArticlePubMedGoogle Scholar
  29. Gouy M, Guindon S, Gascuel O: SeaView version 4: a multiplatform graphical user interface for sequence alignment and phylogenetic tree building. Mol Biol Evol. 2010, 27: 221-224. 10.1093/molbev/msp259.View ArticlePubMedGoogle Scholar
  30. Posada D: ModelTest: phylogenetic model averaging. Mol Biol Evol. 2008, 25: 1253-1256. 10.1093/molbev/msn083.View ArticlePubMedGoogle Scholar
  31. Erixon P, Svennblad B, Britton T, Oxelman B: Reliability of Bayesian posterior probabilities and bootstrap frequencies in phylogenetics. Syst Biol. 2003, 52: 665-673. 10.1080/10635150390235485.View ArticlePubMedGoogle Scholar
  32. Stamatakis A: RAxML-VI-HPC: maximum likelihood-based phylogenetic analyses with thousands of taxa and mixed models. Bioinformatics. 2006, 22: 2688-2690. 10.1093/bioinformatics/btl446.View ArticlePubMedGoogle Scholar
  33. Felsenstein J: Confidence limits on phylogenies: an approach using the bootstrap. Evolution. 1985, 39: 783-791. 10.2307/2408678.View ArticleGoogle Scholar
  34. Ronquist F, Huelsenbeck JP: MRBAYES 3: Bayesian phylogenetic inference under mixed models. Bioinformatics. 2003, 19: 1572-1574. 10.1093/bioinformatics/btg180.View ArticlePubMedGoogle Scholar
  35. Degnan JH: Probabilities of gene tree topologies with intraspecific sampling given a species tree. Estimating Species Trees: Practical and Theoretical Aspects. Edited by: Knowles LL, Kubatko LS. 2010, Hoboken, NJ: Wiley-Blackwell, 53-55.Google Scholar
  36. Avila ML, Tekiel V, Moretti G, Nicosia S, Bua J, Lammel EM, Stroppa MM, Burgos NMG, Sánchez DO: Gene discovery in Triatoma infestans. Parasit Vectors. 2011, 4: 39-10.1186/1756-3305-4-39.PubMed CentralView ArticlePubMedGoogle Scholar
  37. Knowles LL, Kubatko LS: Estimating species trees: an introduction to concepts and models. Estimating Species Trees: Practical and Theoretical Aspects. Edited by: Knowles LL, Kubatko LS. 2010, Hoboken, NJ: Wiley-Blackwell, 1-14.Google Scholar
  38. Neiva A, Pinto C: Estado actual dos conhecimentos sôbre o gênero Rhodnius Stål, com a descripção de uma nova especie. Brasil Med. 1923, 37: 20-24.Google Scholar
  39. Wiens JJ, Tiu J: Highly incomplete taxa can rescue phylogenetic analyses from the negative impacts of limited taxon sampling. PLoS ONE. 2012, 7: e42925-10.1371/journal.pone.0042925.PubMed CentralView ArticlePubMedGoogle Scholar
  40. Dujardin JP, Costa J, Bustamante D, Jaramillo N, Catalá S: Deciphering morphology in Triatominae: the evolutionary signals. Acta Trop. 2009, 110: 101-111. 10.1016/j.actatropica.2008.09.026.View ArticlePubMedGoogle Scholar
  41. Bargues MD, Marcilla A, Dujardin JP, Mas-Coma S: Triatomine vectors of Trypanosoma cruzi: a molecular perspective based on nuclear ribosomal DNA markers. Trans R Soc Trop Med Hyg. 2002, 96 (Suppl 1): S159-S164.View ArticlePubMedGoogle Scholar
  42. De Paula AS, Diotaiuti L, Schofield CJ: Testing the sister-group relationship of the Rhodniini and Triatomini (Insecta: Hemiptera: Reduviidae: Triatominae). Mol Phylogenet Evol. 2005, 35: 712-718. 10.1016/j.ympev.2005.03.003.View ArticlePubMedGoogle Scholar
  43. Patterson JS, Barbosa SE, Feliciangeli MD: On the genus Panstrongylus Berg 1879: evolution, ecology and epidemiological significance. Acta Trop. 2009, 110: 187-199. 10.1016/j.actatropica.2008.09.008.View ArticlePubMedGoogle Scholar
  44. Morrone JJ: Biogeographic areas and transition zones of Latin America and the Caribbean islands based on panbiogeographic and cladistic analyses of the entomofauna. Ann Rev Entomol. 2006, 51: 467-494. 10.1146/annurev.ento.50.071803.130447.View ArticleGoogle Scholar
  45. Fernandez-Loayza R: Triatoma matsunoi nueva especie del norte peruano (Hemiptera, Reduviidae: Triatominae). Rev Per Entomol. 1989, 31: 21-24.Google Scholar
  46. Jurberg J, Galvão C: Hermanlentia n. gen. da tribo Triatomini, com um rol de espécies de Triatominae (Hemiptera, Reduviidae). Mem I Oswaldo Cruz. 1997, 92: 181-185.Google Scholar
  47. Gorla DE, Dujardin JP, Schofield CJ: Biosystematics of Old World Triatominae. Acta Trop. 1997, 63: 127-140. 10.1016/S0001-706X(97)87188-4.View ArticlePubMedGoogle Scholar
  48. Dorn PL, Calderon C, Melgar S, Moguel B, Solorzano E, Dumonteli E, Rodas A, de la Rua N, Garnica R, Monroy C: Two distinct Triatoma dimidiata (Latreille, 1811) taxa are found in sympatry in Guatemala and Mexico. PLoS Negl Trop Dis. 2009, 3: e393-10.1371/journal.pntd.0000393.PubMed CentralView ArticlePubMedGoogle Scholar
  49. Espinoza B, Martínez-Ibarra JA, Villalobos G, De La Torre P, Laclette JP, Martínez-Hernández F: Genetic variation of North American Triatomines (Insecta: Hemiptera: Reduviidae): initial divergence between species and populations of Chagas disease vector. Am J Trop Med Hyg. 2013, 88: 275-284. 10.4269/ajtmh.2012.12-0105.PubMed CentralView ArticlePubMedGoogle Scholar
  50. Alevi KC, Mendonça PP, Pereira NP, Rosa JA, Oliveira MT: Karyotype of Triatoma melanocephala Neiva and Pinto (1923). Does this species fit in the Brasiliensis subcomplex?. Infect Genet Evol. 2012, 12: 1652-1653. 10.1016/j.meegid.2012.06.011.View ArticlePubMedGoogle Scholar
  51. Monteiro FA, Donnelly MJ, Beard CB, Costa J: Nested clade and phylogeographic analyses of the Chagas disease vector Triatoma brasiliensis in Northeast Brazil. Mol Phylogenet Evol. 2004, 32: 46-56. 10.1016/j.ympev.2003.12.011.View ArticlePubMedGoogle Scholar
  52. Dias JCP, Silveira AC, Schofield CJ: The impact of Chagas disease control in Latin America - a review. Mem I Oswaldo Cruz. 2002, 97: 603-612. 10.1590/S0074-02762002000500002.View ArticleGoogle Scholar
  53. Gaunt MW, Miles MA: The ecotopes and evolution of triatomine bugs (Triatominae) and their associated trypanosomes. Mem I Oswaldo Cruz. 2000, 95: 557-565.View ArticleGoogle Scholar

Copyright

© Justi et al.; licensee BioMed Central Ltd. 2014

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Advertisement